Localization of PML I or PML Ib and ICP0 in HSV-1 infected cells.

<p>(A and B). B104-1-1 cells were stably transfected with PMLIb and exposed to 0.1 PFU/cell HSV-1(F). Cells were fixed 28h post infection. Dual color immunofluorescence staining was performed with PG-M3 (diluted 1∶150) (red), and anti-ICP0 antibody (green). Chromosomal DNA was counterstained with cytox orange. Viral proteins were not detected by the PG-M3 antibody in HSV-1 infected Vector control cells (A), and ICP0 stained uniquely from PML in HSV-1 infected PML I+PML Ib co-transfected cells (compare the heavily infected cell in upper right to the recently infected cell in the lower left corner) (B). (C and D) B104-1-1 stably transfected with PML I (C) or PML Ib (D) were exposed to 0.1 PFU/cell HSV-1 (F). Cells were fixed at 2h and 28h post infection as indicated. Images were captured with a Zeiss 510 LSM confocal microscope. (E) Further analysis using the LSM Image Browser software demonstrates co-localization of PML Ib and ICP0 in the cytoplasm of a PML Ib expressing cell infected for 28 h with 0.1 PFU/cell. Red and green represent PML Ib and ICP0 respectively, while cytox orange staining is represented by blue in both the upper image and intensity profile (bottom). The intensity profile was software generated from the fluorescence intensity values encompassed along the red line (upper image).